ARTP诱变技术选育高产谷氨酰胺酶的曲霉菌种Selection and Breeding of Aspergillus oryzae with High-yield Glutaminase by ARTP Induced Mutation Technique
周其洋,张书泰
摘要(Abstract):
为了选育高产谷氨酰胺酶的突变菌株,选择综合性能优良的酱油生产曲霉菌株A1428,利用常压室温等离子体(ARTP)诱变育种技术,经平板初筛和酱油制曲、发酵复筛,结合96孔板法建立高通量筛选诱变菌株的方法,得到一株高产谷氨酰胺酶菌株A380。结果表明:ARTP诱变条件为功率120W,气流量10L/min,诱变时间120s,致死率达90%。菌株A380谷氨酰胺酶活力为10U/g,较出发菌株高25%。发酵原油谷氨酸含量为12g/L,提高了20%。同时,诱变菌株A380原油中鲜味氨基酸、甜味氨基酸较出发菌株A1428都有一定的提升,苦味氨基酸相对较低,具有进一步研究其工业化生产的价值。
关键词(KeyWords): ARTP;谷氨酰胺酶;菌种选育;高通量筛选
基金项目(Foundation): 广东省调味食品先进制造工程技术研究中心(gc001886);; 2017年度广东省科技发展专项资金项目公益研究与能力建设类广东省调味食品生物发酵先进技术企业重点实验室建设项目(2017B030302002)
作者(Author): 周其洋,张书泰
参考文献(References):
- [1]周池虹伶.一株解淀粉芽孢杆菌产谷氨酰胺酶的应用研究[D].广州:华南理工大学,2016:14-35.
- [2]邹谋勇,朱新贵.酱油发酵醪中谷氨酰胺酶活性蛋白序列分析[J].食品科学,2017,38(22):143-148.
- [3]李鹏,张艳芳,王选年.米曲霉双菌株组合制曲对产蛋白酶的影响[J].食品与生物技术学报,2018,37(2):206-210.
- [4]Sathish T,Lakshmi G S,Rao C S,et al.Mixture design as first step for improved glutaminase production in solid-state fermentation by isolated Bacillus[J].Letters in Applied Microbiology,2010,47(4):256-262.
- [5]Yano T,Ito M,Tomita K,et al.Purification and properties of glutaminase fromAspergillus oryzae[J].Journal of Fermentation Technology,1988,66(2):137-143.
- [6]Ito K,Koyama Y,Hanya Y.Identification of the glutaminase genes of Aspergillus sojae involved in glutamate production during soy sauce fermentation[J].Bioscience Biotechnology&Biochemistry,2013,77(9):1832-1840.
- [7]Ueki T,Noda Y,Teramoto Y,et al.Increasing glutaminase activity in shoyu koji using a mixed culture of two koji moulds[J].World Journal of Microbiology&Biotechnology,1994,10(6):694-696.
- [8]周斌,王静,周其洋,等.米曲霉基因组与酶系表达研究进展[J].食品与发酵工业,2018,44(6):293-299.
- [9]陆祥,何维.一株产谷氨酰胺酶菌种的筛选及产酶分析[J].轻工科技,2017,33(5):5-6.
- [10]Xia J.UV and 60Co-γ-ray combined mutagenesis of Aspergillus niger for the enhancement of glucoamylase activity[J].Mycosystema,2014.
- [11]李鹏,张艳芳,王军,等.紫外诱变选育米曲霉氨肽酶高产菌株[J].中国调味品,2017,42(1):61-64.
- [12]叶菁,蒋雪薇,罗晓明,等.高蛋白酶酶活酱油发酵用米曲霉的复合诱变选育[J].食品与机械,2017,33(1):7-10.
- [13]舒冬梅,王德良,宋绪磊,等.常压室温等离子体诱变(ARTP)及高通量筛选高产蛋白酶米曲霉的初探[J].中国调味品,2016,41(12):67-73.
- [14]陈锡剑,陈建华.常压室温等离子体诱变选育高产曲酸米曲霉[J].化学与生物工程,2015,32(5):52-54.
- [15]蒋莹.谷氨酰胺转氨酶产生菌的常压室温等离子体诱变选育[D].上海:华东理工大学,2017:49-56.
- [16]Zhao G,Hou L,Lu M,et al.Construction of the mutant strain in Aspergillus oryzae 3.042for abundant proteinase production by the N+ion implantation mutagenesis[J].International Journal of Food Science&Technology,2012,47(3):504-510.
- [17]童星,彭勃.一株高酶活力米曲霉菌株的选育及其在酱油生产中的应用[J].中国酿造,2018,37(11):51-55.
- [18]文志州.中心组合设计在优化发酵酱种曲培养基中的应用研究[J].中国调味品,2016,41(4):117-121.
- [19]李荔,童星.酱油酿造中米曲霉和酱油曲霉复合制曲的研究[J].中国调味品,2018,43(5):145-148.
- [20]谭永水,郭琳,刘新利,等.三种酱油发酵曲霉酶活力的比较[J].中国调味品,2017,42(11):24-28.
- [21]Cui Chun,Zhao Mouming,Li Dan,et al.Biochemical changes of traditional Chinese type soy sauce produced in four seasons during processing[J].CyTA-Journal of Food,2014,12(2):166-175.
- [22]Zhang Liqiang,Zhou Rongqing,Cui Ruiying,et al.Characterizing soy sauce moromi manufactured by high-salt dilute-state and low-salt solid-state fermentation using multiphase analyzing methods[J].Journal of Food Science,2016,81(11):1-8.